Note: the data displayed was generated using our legacy chemistry (SQK-LSK109), but the outcomes and recommendations are still applicable to our current chemistry.

The final stage of DNA library preparation for the Ligation Sequencing Kit is to elute the library in Elution Buffer. Typically, after the elution step, the library is mixed with the sequencing buffer and loading beads, then immediately loaded onto the flow cell. However, users may want to store the library in Elution Buffer and load the flow cell at a later point in time.

To demonstrate the stability of the sequencing libraries produced using the Ligation Sequencing Kit, a large-scale library was generated using Lambda DNA as the template. The library was aliquoted into single-use tubes and stored under different conditons for various lengths of time.

No significant decrease in output or read length was observed when storing the libraries at -80°C, 4°C or 21°C over the course of the 12 week study. However, storage at -20°C for 12 weeks and 30°C for 1 week led to a decrease in output. Therefore, we recommend storing libraries in Eppendorf DNA LoBind tubes at 4°C for short-term or repeated use. For single use or longer-term storage of more than 3 months, we recommend storing at -80°C.

Figure 1. The output (Mb/ch/h) and read N50 values for libraries that have been stored at various temperatures for varying lengths of time. Sequencing libraries were generated using SQK-LSK109 and stored in Elution Buffer for varying lengths of time. After storage, the libraries were mixed with sequencing buffer and loading beads before loading onto a MinION flow cell for sequencing.

Version	Changelog
v2, May 2025	Added wording to clarify results are applicable to newer kit chemistry
v1, August 2020	Initial protocol publication